Successful expression of the Bordetella petrii nitrile hydratase activator P14K and the unnecessary role of Ser115 View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2016-12

AUTHORS

Weifeng Sun, Longbao Zhu, Xianggui Chen, Ping Chen, Lingling Yang, Wenwu Ding, Zhemin Zhou, Yi Liu

ABSTRACT

BACKGROUND: The activator P14K is necessary for the activation of nitrile hydratase (NHase). However, it is hard to be expressed heterogeneously. Although an N-terminal strep tagged P14K could be successfully expressed from Pseudomonas putida, various strategies for the over-expression of P14K are needed to facilitate further application of NHase. RESULTS: P14K was successfully expressed through fusing a his tag (his-P14K), and was over-expressed through fusing a gst tag (gst-P14K) at its N-terminus in the NHase of Bordetella petrii DSM 12804. The stability of gst-P14K was demonstrated to be higher than that of the his-P14K. In addition, the Ser115 in the characteristic motif CXLC-Ser115-C of the active center of NHase was found to be unnecessary for NHase maturation. CONCLUSIONS: Our results are not only useful for the NHase activator expression and the understanding of the role of Ser115 during NHase activation, but also helpful for other proteins with difficulty in heterologous expression. More... »

PAGES

21

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/s12896-016-0252-2

DOI

http://dx.doi.org/10.1186/s12896-016-0252-2

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https://app.dimensions.ai/details/publication/pub.1053539745

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/26897378


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43 schema:description BACKGROUND: The activator P14K is necessary for the activation of nitrile hydratase (NHase). However, it is hard to be expressed heterogeneously. Although an N-terminal strep tagged P14K could be successfully expressed from Pseudomonas putida, various strategies for the over-expression of P14K are needed to facilitate further application of NHase. RESULTS: P14K was successfully expressed through fusing a his tag (his-P14K), and was over-expressed through fusing a gst tag (gst-P14K) at its N-terminus in the NHase of Bordetella petrii DSM 12804. The stability of gst-P14K was demonstrated to be higher than that of the his-P14K. In addition, the Ser115 in the characteristic motif CXLC-Ser115-C of the active center of NHase was found to be unnecessary for NHase maturation. CONCLUSIONS: Our results are not only useful for the NHase activator expression and the understanding of the role of Ser115 during NHase activation, but also helpful for other proteins with difficulty in heterologous expression.
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